U.S. patent application number 11/383496 was filed with the patent office on 2008-01-03 for the production of high-activity superoxide dismutase (sod) and application in methods of both the solid-state and liquid-state fermentation.
Invention is credited to Chang-Yu Chuang, Pao-Chuan Hsieh.
Application Number | 20080003641 11/383496 |
Document ID | / |
Family ID | 38877133 |
Filed Date | 2008-01-03 |
United States Patent
Application |
20080003641 |
Kind Code |
A1 |
Hsieh; Pao-Chuan ; et
al. |
January 3, 2008 |
THE PRODUCTION OF HIGH-ACTIVITY SUPEROXIDE DISMUTASE (SOD) AND
APPLICATION IN METHODS OF BOTH THE SOLID-STATE AND LIQUID-STATE
FERMENTATION
Abstract
The present invention of "A production method for high-activity
superoxide dismutase (SOD) and application in methods of both the
solid-state and liquid-state fermentation" lies in processing the
strains of Bacillus subtilis with quantifying the activity of
superoxide dismutase (SOD) in fermentation broth by nitro blue
tetrazolium (NBT) reduction method, and then one kind of
high-activity SOD-producing strain, which will be acquired while
after the dialysis process, named Strain S-8 and afterward, this
strain S-8 will be utilized to produce SOD by performing the
liquid-state and solid-state fermentation methods respectively.
Inventors: |
Hsieh; Pao-Chuan; (Nei Pu
Hsiang, TW) ; Chuang; Chang-Yu; (Nei Pu Hsiang,
TW) |
Correspondence
Address: |
LEONG C LEI
PMB # 1008, 1867 YGNACIO VALLEY ROAD
WALNUT CREEK
CA
94598
US
|
Family ID: |
38877133 |
Appl. No.: |
11/383496 |
Filed: |
May 16, 2006 |
Current U.S.
Class: |
435/69.1 ;
435/192; 435/252.3 |
Current CPC
Class: |
C12N 9/0089
20130101 |
Class at
Publication: |
435/69.1 ;
435/192; 435/252.3 |
International
Class: |
C12N 9/08 20060101
C12N009/08; C12P 21/06 20060101 C12P021/06 |
Claims
1. A kind of high-activity superoxide dismutase (SOD), named Strain
S-8, where the said Strain S-8 has been deposited at the Food
industry Research and Development Institute located in Hsinchu
City, Taiwan, on the date of Feb. 9, 2006 with the Depositary
Serial No. BCRC 910306.
2. A kind of production method for high-activity superoxide
dismutase (SOD) and application in methods of both the solid-state
and liquid-state fermentation lies in filtrating Natto product to
generate the strains of Bacillus Subtilis and then quantifying the
activity of superoxide dismutase (SOD) in fermentation broth by
nitro blue tetrazolium (NBT) reduction method and, in addition, it
is applicable to acquire the high-activity SOD-producing strain,
while after the dialysis process, which is named Strain S-8, and
afterward this strain S-8 will be utilized to produce SOD by
performing the liquid-state and solid-state fermentation methods
respectively.
3. The liquid-state fermentation method as claimed in claim 2,
wherein it is applicable to achieve the nourishment condition of
optimum rapid growth, in which the groups or combinations for
carbon source consisting of glucose, D-sorbitol solution, and
dissoluble starch etc., and the groups or combinations for the
nitrogen source consisting of soybean powder, beef extract, and
peptone etc.
4. The liquid-state fermentation method as claimed in claim 3,
wherein its pH value is about 6.about.7.5.
5. The solid-state fomentation method as claimed in claim 2,
wherein it comprises the combination of wheat acrospire and soybean
together with the additive of sugar in order to perform die
solid-state fermentation process, for fermenting at least 72
hours.
6. The solid-state fermentation method as claimed in claim 5,
wherein it comprises the combination of wheat acrospire and soybean
in size of 10.about.20 mesh diameter.
7. The solid-state fermentation method as claimed in claim 5,
wherein the additive of sugar is the groups or combinations of such
as glucose, D-sorbitol solution, and dissoluble starch, etc.
8. The solid-state fermentation method as claimed in claim 5,
wherein the proportional ratio for sugar additive is 5%.
Description
BACKGROUND OF THE INVENTION
[0001] (a) Technical Field of the Invention
[0002] The present invention relates to the utilization of strain
of Bacillus Subtilis, wherein the dialysis process is performed in
order to acquire the high-activity SOD-producing strain by
performing the liquid-state and solid-state fermentation methods
respectively and, in addition, to study the optimum cultivation
conditions for said strain during the production of SOD.
[0003] (b) Description of the Prior Art
[0004] Reactive Oxygen Species (ROS) comprises free radicals, such
as Superoxide Anion Free Radicals (O2--.), Hydrogen Peroxide (H2O2)
and Hydroxyl Free Radicals (HO.) etc., which are matters generated
unavoidably by the living organisms during oxidative
metabolism.
[0005] In recent years, the studies related to the Reactive Oxygen
Species (ROS) has already been proved that it has a toxicity to
cells and is applicable to activate the mutations of living
organisms. The free radicals can oxidize unsaturated fatty acids on
cell membrane and hence causing a peroxidation reaction, destroying
cell membranes and, in addition, resulting in oxygenation of
protein as well as damages to DNA. In order to prevent from the
attacks by free radicals, thus the living organisms has developed a
kind of defense mechanism, such as non-enzymatic and enzymatic
defense mechanism, wherein the non-enzymatic defense mechanism
comprising matters of iso-ascorbic acid and tocophenol etc. and the
enzymatic defense mechanism comprising matters of superoxide
dismutase (SOD; EC 1.15.1.1), and catalase etc. so as to eliminate
free radicals inside the human body as well as relief the condition
with excessive free radicals.
[0006] Superoxide dismutase (SOD) is a kind of protein having a
metal matter positioned at the activity center, being classified
into three classifications according to metal combined thereat;
Cu/Zn Superoxide dismutase (Cu/Zn-SOD), Fe Superoxide dismutase
(Fe-SOD) and Mn Superoxide dismutase (Mn-SOD), SOD is the earliest
cleaned enzyme while generating the reactive oxygen species (ROS)
inside the human body, where the relevant mechanism is to transform
the superoxide anion fee radicals generating firstly in the
reactive oxygen species (ROS) into hydrogen peroxide (H2O2) and
oxygen molecule, by dismutation in order to relief the accumulation
of excessive reactive oxygen species (ROS) inside the human
body.
[0007] The current studies related to the SOD are mostly focusing
on the reactive units inside the SOD itself, such, as the
comparison of content of superoxide dismutase (SOD) among Taiwanese
medicinal mushrooms, or producing SOD with the fungi--Aspergillus
Oryzae by performing the solid-state fermentation method, so as to
study the activity and yield, however due to the fermentation time
of fungi is much longer than others, thus mere there is very few
seldom to discuss and study in the field of creating SOD activity
with repaid growing germ by the fermentation method.
[0008] Additionally, there are various applications of SOD
currently, for example, SOD has already been used in the clinical
medicine, e.g. in improving patient's autoimmune diseases,
inflammatory disease treatments and relief of muscular fatigue,
regarding to other applications, where the SOD is also applied in
the developments of the functional foods, food fortification and
dietary supplements as well as cosmetics, therefore, if we can find
out the high-yield type SOD-producing strains among the
microorganism, producing the uniform quality products in mass
production with a low cost, thus enhancing the whole economic
benefits while acting as an additive in industries of health foods,
cosmetics and pharmaceutics.
SUMMARY OF THE INVENTION
[0009] The present invention of "A production method for
high-activity superoxide dismutase (SOD) and application in methods
of both the solid-state and liquid-state fermentation" lies in
providing the mass production of Bacillus Subtilis in order to
acquire the high-activity SOD-producing strain, named Strain S-8,
which has been deposited at the Food Industry Research and
Development Institute located in Hsinchu City, Taiwan, on the date
of Feb. 9, 2006 with the Depositary Serial No. BCRC 910306.
[0010] The present invention of "A production method for
high-activity superoxide dismutase (SOD) and application in methods
of both the solid-state and liquid-state fermentation" lies in
performing the dialysis process in order to acquire the
high-activity SOD-producing strain by performing the liquid-state
and solid-state fermentation methods respectively and, in addition,
to provide the optimum cultivation conditions for the said strain
during the production of SOD.
[0011] The foregoing object and summary provide only a brief
introduction to the present invention. To fully appreciate these
and other objects of the present invention as well as the invention
itself, all of which will become apparent to those skilled in the
art, the following detailed description of the invention and the
claims should be read in conjunction with the accompanying
drawings. Throughout the specification and drawings identical
reference numerals refer to identical or similar parts.
[0012] Many other advantages and features of the present invention
will become manifest to those versed in the art upon making
reference to the detailed description and the accompanying sheets
of drawings in which a preferred structural embodiment
incorporating the principles of the present invention is shown by
way of illustrative example.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] The present invention will be apparent to those skilled in
the art by reading the following detailed description of a
preferred embodiment thereof, with reference to the attached
drawings, in which;
[0014] FIG. 1 illustrates variation related to SOD activity, pH
value and total bacterial counts for the strain of Bacillus
Subtilis S-8 during the process of liquid-state fermentation;
[0015] FIG. 2 illustrates variation related to SOD activity, pH
value and total bacterial counts for the strain of Bacillus
Subtilis S-8 during the process of solid-state fermentation.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0016] The following descriptions are of exemplary embodiments
only, and are not intended to limit the scope, applicability or
configuration of the invention in any way. Rather, the following
description provides a convenient illustration for implementing
exemplary embodiments of the invention. Various changes to the
described embodiments may be made in the function and arrangement
of the elements described without departing from the scope of the
invention as set forth in the appended claims.
[0017] The present invention relates to a kind of production method
for high-activity superoxide dismutase (SOD) and application in
methods of both the solid-state and liquid-state fermentation lies
in filtrating Natto product bought from the local markets to
generate the 270 strains of Bacillus Subtilis and then quantifying
the activity of superoxide dismutase (SOD) in fermentation broth by
nitro blue tetrazolium (NBT) reduction method and, in addition, it
is applicable to acquire the high-activity SOD-producing strain,
while after the dialysis process, which, is named Strain S-8, and
afterward this strain S-8 will be utilized to produce SOD by
performing the liquid-state and solid-state fermentation methods
respectively.
[0018] Without the original studying in nourishment condition, the
Strain S-8 is capable to produce the SOD with an activity of 153
U/mL, and it is applicable to achieve the nourishment condition of
optimum rapid growth by performing the liquid-state fermentation
method, in which the groups or combinations for carbon source
consisting of glucose, D-sorbitol solution, and dissoluble starch,
and the groups or combinations for the nitrogen source consisting
of soybean powder, beef extract, and peptone, with a pH value of
about 6.about.7.5, more over, the Strain S-8 can produce the
superoxide dismutase (SOD) with an improved activity of 2600
U/mL.
[0019] The production capability of Strain S-8 is performed via an
experiment by the solid-state fermentation method, wherein
solid-state fermentation substrate comprising the combination of
wheat acrospire in 10.about.20 mesh diameter and soybean together
with the additive of 5% sugar, such as the groups or combinations
of glucose, D-sorbitol solution, dissoluble starch, in order to
perform the solid-state fermentation process, for fermenting at
least 72 hours, wherein the activity of superoxide dismutase (SOD)
contained in fermentation product can reach up to 21,000 U/g, which
is much higher than the control group with 645 U/g which didn't
have any Strain S-8 at all.
[0020] Although the present invention has been described with
reference to the preferred embodiment thereof, it is apparent to
those skilled in the art that a variety of modifications and
changes may be made without departing from the scope of the present
invention which is intended to be defined by the appended
claims.
[0021] It will be understood that each of the elements described
above, or two or more together may also find a useful application
in other types of methods differing from the type described
above.
[0022] While certain novel features of this invention have been
shown and described and are pointed out in the annexed claim, it is
not intended to be limited to the details above, since it will be
understood that various omissions, modifications, substitutions and
changes in the forms and details of the device illustrated and in
its operation can be made by those skilled in the art without
departing in any way from the spirit of the present invention.
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